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. 2015 Apr 23;33(5):1390–1404. doi: 10.1002/stem.1926

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© 2014 The Authors. STEM CELLS published by Wiley Periodicals, Inc. on behalf of AlphaMed Press

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Retinoic acid (RA) signalling in EpiSC reprogramming. (A): RA signalling as measured by a luciferase assay in EpiSCs cultured in N2B27/activin/FGF2 with VA and other retinoids. The activities are shown relative to that of Renilla luciferase and normalized to that of EpiSCs in −VA. (B): A diagram of EpiSC reprogramming in 2i/LIF. (C): The effects of retinoids on reprogramming Oct4‐GFP reporter EpiSCs to induced pluripotent stem cells by LRH1. *, p < .05; **, p < .01. Scale bar = 200 µm. (D): RA signalling at various stages of EpiSC reprogramming. **, p < .01. (E): The effects of expressing RARG (under the CMV early enhancer/chicken β actin promoter) on reprogramming EpiSCs by LRH1. Note that even small amounts of Rarg PB transposon have deleterious effects. *, p < .05; **, p < .01. (F): Blocking LRH1‐mediated EpiSC reprogramming by ligand depletion (citral), RA receptor inhibition (CD2665) or expression of RARA‐DN. Reprogramming was partially restored by 0.1 nM ATRA. Oct4‐GFP⁺ colonies were scored on day 9 after transfection. *, p < .05; **, p < .01. In all the experiments, Oct4‐GFP⁺ colonies were scored on day 9 after transfection. The experiments were repeated at least three times, and the error bars show standard deviations from the mean of triplicate determinations in one representative experiment. Abbreviations: ATRA, all‐trans retinoic acid; 9cRA, 9‐cis‐retinoic acid; EpiSCs, epiblast stem cells; FGF2, fibroblast growth factor 2; LIF, leukemia inhibitory factor; LRH1, liver receptor homolog‐1; RARA‐DN, dominant‐negative form of RARA; RARG, retinoic acid receptor gamma; VA, vitamin A.